CN103721136B - A kind of preparation method of Yanyan slice and application - Google Patents
A kind of preparation method of Yanyan slice and application Download PDFInfo
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- 239000010231 banlangen Substances 0.000 claims abstract description 11
- 230000004663 cell proliferation Effects 0.000 claims abstract description 11
- 210000000582 semen Anatomy 0.000 claims abstract description 11
- 206010020880 Hypertrophy Diseases 0.000 claims abstract description 9
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 9
- 201000011510 cancer Diseases 0.000 claims abstract description 9
- 230000001629 suppression Effects 0.000 claims abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 27
- 238000000605 extraction Methods 0.000 claims description 9
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- 238000002474 experimental method Methods 0.000 description 8
- 238000000034 method Methods 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 210000003800 pharynx Anatomy 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
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- 239000006228 supernatant Substances 0.000 description 2
- 235000019640 taste Nutrition 0.000 description 2
- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
- QZKDXSOKZHLFHE-UHFFFAOYSA-N 4-(1,3-benzodioxol-5-yl)benzoic acid Chemical compound C1=CC(C(=O)O)=CC=C1C1=CC=C(OCO2)C2=C1 QZKDXSOKZHLFHE-UHFFFAOYSA-N 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
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- 201000007100 Pharyngitis Diseases 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 241001530126 Scrophularia Species 0.000 description 1
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- 239000002250 absorbent Substances 0.000 description 1
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- 230000000954 anitussive effect Effects 0.000 description 1
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- ZZVUWRFHKOJYTH-UHFFFAOYSA-N diphenhydramine Chemical compound C=1C=CC=CC=1C(OCCN(C)C)C1=CC=CC=C1 ZZVUWRFHKOJYTH-UHFFFAOYSA-N 0.000 description 1
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a kind of preparation method of Yanyan slice, by Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g make as crude drug, employing supercritical extraction is prepared from, content is improved a lot, dose reduces, and present invention also offers the application of Yanyan slice in preparation suppression mouse hypertrophy cell cancer P815 cell proliferation.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of preparation method and application of Yanyan slice.
Background technology
Yanyan slice is recorded in Ministry of Public Health standard WS3-B-0331-90, prescription be Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g, above 12 tastes, except Oleum menthae, Flos Farfarae is ground into fine powder, sieves; Ten tastes such as all the other Radix Scrophulariaes decoct with water secondary, 3 hours first times, second time 2 hours, and collecting decoction leaves standstill 24 hours, get supernatant, be condensed into thick paste, mix with Common Coltsfoot Flower, dry, make granule, spray is with Oleum menthae, and mixing, is pressed into 1000, sugar coating, obtains final product.Energy nourishing YIN and moistening the lung, heat-clearing and toxic substances removing, clearing throat, antitussive is antipruritic.For the dry pharynx that chronic pharyngitis causes, itching throat, the diseases such as irritable cough.
In prior art, not yet there is Yanyan slice extracting the report adopting supercritical technology in preparation, and adopt soak by water and the method for beating powder, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of preparation method of Yanyan slice.
Another object of the present invention is to provide a kind of Yanyan slice to suppress the application in mouse hypertrophy cell cancer P815 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of Yanyan slice, by Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g makes as crude drug, described method is made up of the following step: get Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g joins in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.25g of every sheet.
The preparation method of above-mentioned a kind of Yanyan slice, described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of above-mentioned a kind of Yanyan slice, described CO
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned a kind of Yanyan slice suppresses the application in mouse hypertrophy cell cancer P815 cell proliferation in preparation, Yanyan slice is by Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g makes as crude drug, preparation method is made up of the following step: get Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.25g of every sheet.
Above-mentioned a kind of Yanyan slice suppresses the application in mouse hypertrophy cell cancer P815 cell proliferation in preparation, and the percent by volume that the entrainer of CO2 supercritical extraction described in the preparation method of Yanyan slice accounts for total extractant is 5%.
Above-mentioned a kind of Yanyan slice suppresses the application in mouse hypertrophy cell cancer P815 cell proliferation in preparation, the extracting pressure 20MPa of CO2 supercritical extraction described in the preparation method of Yanyan slice, temperature 40 DEG C, CO2 flow 2ml/g crude drug min, extraction time 160min.
In prior art, the every sheet 0.25g of Yanyan slice, each 5,3 times on the one, adopt the every sheet 0.25g of Yanyan slice that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only need 3 at every turn, within 1st, take 3 times, under the condition with more active component, greatly reduce dose.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 30 DEG C, CO
2flow 1m1/g crude drug min, extraction time 150min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.25g of every sheet.
Embodiment 2
Get Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 30MPa, temperature 50 C, CO
2flow 3m1/g crude drug min, extraction time 180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.25g of every sheet.
Embodiment 3
Get Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, temperature 40 DEG C, CO
2flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.25g of every sheet.
Embodiment 4: Yanyan slice suppresses the experimentation data of P815 cell proliferation
1 experiment material
1.1 experiment cell strains
Mouse hypertrophy cell cancer P815 cell, Nanjing Medical University's laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: Yanyan slice of the present invention: prepare by embodiment 3 method.
Medicinal liquid liquid storage: take 100mg Yanyan slice, is dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ ldoff pipe subpackages ,-20 DEG C of storages, and simultaneously 0.2 μm of frit dehydrated alcohol is in order to the use of matched group.
1.3 experiment reagent
DMEM(GIBCO company Cat.No.12100-061Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3(Shanghai hundred million chemical reagent company limited Cat.No.11810-033Lot.No.1088387 of a specified duration); Trypsin(AMRESCO company lot number: 2010/04); EDTA(AMRESCO company lot number: 2009/10); PenicillinGSodiumSalt(AMRESCO company lot number: 2010242); StreptomycinSulfate(AMRESCO company lot number: 2010382); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); PBS(laboratory autogamy);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (the accurate experimental facilities company in Shanghai model: DHG9123A); Refrigerator (Siemens Company's model: KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000); 0.2 μm of filter (MILLIPORE model: SLGP033RB); 10cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2 experimental techniques
1) P815 cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out cellar culture (10cm culture dish), when Growth of Cells is to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 DEG C of digestion 2min, add 5ml complete medium neutralization reaction wherein, proceeded in centrifuge tube after piping and druming cell, the centrifugal 5min of 1000rpm, adjustment concentration of cell suspension 3 × 104/ml.
2) enter in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, and culture plate puts into cell culture incubator (37 DEG C, 5%CO2) cellar culture.
3) according to cell growth status, generally grow to 50%-70%, add Yanyan slice solution, continue to cultivate 24h.
4) add 20 μ lMTT solution (5mg/ml, i.e. 0.5%MTT) after 24h, continue to cultivate 4h.
5) after 4h, buckle method removes supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm place.
6) background (do not add cell, only add culture fluid) is set, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide) simultaneously, often organizes the multiple hole of setting 6.
7) result represents with the suppression ratio of medicine to cell:
Cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment repetition 3 times.
3 statistical dispositions
Adopt the correlation analysis in MicrosoftExcel2003 software and Studentt inspection, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to P815 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has pole significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 Yanyan slice is to P815 cell inhibitory effect influence research
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Yanyan slice can suppress P815 cell proliferation, and reduce the Growth of Cells number of P815 cell, this effect is dose dependent.
Claims (3)
1. the application of Yanyan slice in preparation suppression mouse hypertrophy cell cancer P815 cell proliferation, it is characterized in that Yanyan slice is by Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g makes as crude drug, preparation method is made up of the following step: get Radix Scrophulariae 120g, Radix Stemonae 90g, Radix Asparagi 90g, Cortex Moutan 90g, Radix Ophiopogonis 90g, Flos Farfarae 90g, Semen Oroxyli 30g, Radix Rehmanniae 90g, Radix Isatidis 150g, Fructus Canarii 90g, Periostracum Cicadae 30g, Oleum menthae 0.3g, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.25g of every sheet.
2. a kind of Yanyan slice suppresses the application in mouse hypertrophy cell cancer P815 cell proliferation in preparation according to claim 1, CO described in the preparation method that it is characterized in that Yanyan slice
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of Yanyan slice suppresses the application in mouse hypertrophy cell cancer P815 cell proliferation in preparation according to claim 1, CO described in the preparation method that it is characterized in that Yanyan slice
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
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Effective date of registration: 20191206 Address after: Room 306, building 3, business center, No. 82, Huayuan West Road, Chuzhou, Anhui Patentee after: CHUZHOU TIANSHUN PRODUCTIVITY PROMOTION Co.,Ltd. Address before: 226500 Yang Zhuangju, Dong Chen Town, Rugao City, Jiangsu, Nantong Patentee before: RUGAO DINGBEI Hospital |
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Effective date of registration: 20221223 Address after: 276000 No. 95, Huaxia Road, economic and Technological Development Zone, Linyi City, Shandong Province Patentee after: SHANDONG TIANCHENG STEEL STRUCTURE Co.,Ltd. Address before: 239000 incubation workshop, room 306, building 3, entrepreneurship center, No.82 Huayuan West Road, Chuzhou City, Anhui Province Patentee before: CHUZHOU TIANSHUN PRODUCTIVITY PROMOTION Co.,Ltd. |